Rapid development of sensitive, high-throughput, quantitative and highly selective mass spectrometric targeted immunoassays for clinically important proteins in human plasma and serum

Bryan Krastins, Amol Prakash, David A. Sarracino, Dobrin Nedelkov, Eric E. Niederkofler, Urban A. Kiernan, Randall Nelson, Maryann S. Vogelsang, Gouri Vadali, Alejandra Garces, Jennifer N. Sutton, Scott Peterman, Gregory Byram, Bruno Darbouret, Joëlle R. Pérusse, Nabil G. Seidah, Benoit Coulombe, Johan Gobom, Erik Portelius, Josef PanneeKaj Blennow, Vathany Kulasingam, Lewis Couchman, Caje Moniz, Mary F. Lopez

Research output: Contribution to journalArticle

77 Scopus citations

Abstract

Objectives: The aim of this study was to develop high-throughput, quantitative and highly selective mass spectrometric, targeted immunoassays for clinically important proteins in human plasma or serum. Design and methods: The described method coupled mass spectrometric immunoassay (MSIA), a previously developed technique for immunoenrichment on a monolithic microcolumn activated with an anti-protein antibody and fixed in a pipette tip, to selected reaction monitoring (SRM) detection and accurate quantification of targeted peptides, including clinically relevant sequence or truncated variants. Results: In this report, we demonstrate the rapid development of MSIA-SRM assays for sixteen different target proteins spanning seven different clinically important areas (including neurological, Alzheimer's, cardiovascular, endocrine function, cancer and other diseases) and ranging in concentration from pg/mL to mg/mL. The reported MSIA-SRM assays demonstrated high sensitivity (within published clinical ranges), precision, robustness and high-throughput as well as specific detection of clinically relevant isoforms for many of the target proteins. Most of the assays were tested with bona-fide clinical samples.In addition, positive correlations, (R2 0.67-0.87, depending on the target peptide), were demonstrated for MSIA-SRM assay data with clinical analyzer measurements of parathyroid hormone (PTH) and insulin growth factor 1 (IGF1) in clinical sample cohorts. Conclusions: We have presented a practical and scalable method for rapid development and deployment of MS-based SRM assays for clinically relevant proteins and measured levels of the target analytes in bona fide clinical samples. The method permits the specific quantification of individual protein isoforms and addresses the difficult problem of protein heterogeneity in clinical proteomics applications.

Original languageEnglish (US)
Pages (from-to)399-410
Number of pages12
JournalClinical Biochemistry
Volume46
Issue number6
DOIs
StatePublished - Apr 2013

Keywords

  • Biomarker
  • Clinical proteomics
  • Immunoassay
  • Mass spectrometry
  • Proteomics
  • SRM assay

ASJC Scopus subject areas

  • Clinical Biochemistry

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    Krastins, B., Prakash, A., Sarracino, D. A., Nedelkov, D., Niederkofler, E. E., Kiernan, U. A., Nelson, R., Vogelsang, M. S., Vadali, G., Garces, A., Sutton, J. N., Peterman, S., Byram, G., Darbouret, B., Pérusse, J. R., Seidah, N. G., Coulombe, B., Gobom, J., Portelius, E., ... Lopez, M. F. (2013). Rapid development of sensitive, high-throughput, quantitative and highly selective mass spectrometric targeted immunoassays for clinically important proteins in human plasma and serum. Clinical Biochemistry, 46(6), 399-410. https://doi.org/10.1016/j.clinbiochem.2012.12.019